REVIEWS

STRESS

Epigenetic and transgenerational

reprogramming of brain development
Tracy L. Bale
Abstract | Neurodevelopmental programming — the implementation of the genetic and
epigenetic blueprints that guide and coordinate normal brain development — requires tight
regulation of transcriptional processes. During prenatal and postnatal time periods,
epigenetic processes fine-tune neurodevelopment towards an end product that determines
how an organism interacts with and responds to exposures and experiences throughout life.
Epigenetic processes also have the ability to reprogramme the epigenome in response to
environmental challenges, such as maternal stress, making the organism more or less
adaptive depending on the future challenges presented. Epigenetic marks generated within
germ cells as a result of environmental influences throughout life can also shape future
generations long before conception occurs.

Department of Biomedical
Sciences, School of Veterinary
Medicine and Department of
Psychiatry, Perelman School
of Medicine, University of
Pennsylvania, Philadelphia,
Pennsylvania 19104, USA.
e-mail: tbale@vet.upenn.edu
doi:10.1038/nrn3818
Published online 29 April 2015
Environmental changes that perturb an animal’s home-
ostasis — including stress, nutritional challenges and
infection — can influence the developing and matur-
ing brain and thereby alter the original blueprint and
establish a new one in a process known as reprogram-
ming 1. There is a great history of studies that have
examined this process, and it has recently become clear
that epigenetic mechanisms, including DNA meth-
ylation, histone modification and the actions of small
non-coding RNAs, have an important role in such
neuro­developmental reprogramming 2–5. However, the
precise mechanisms by which exposure to environmen-
tal perturbations can influence long-term outcomes are
highly complex and depend on the type and the severity
of the exposure, whether it occurs during a window of
vulnerability in the target circuit or system, the organ-
ismal sex and whether there are epigenetic changes in
germ cells (FIG. 1).
The timing of an environmental exposure or experi-
ence may determine whether the event has limited and
specific epigenetic effects or whether it more broadly
affects the epigenome (BOX 1) as well as how long the
effect persists (acutely or across generations). As epige-
netic processes are integrally involved in neurodevel-
opment and maturation, insults that are experienced
during these key periods (which roughly correspond
to early or late gestation) would be predicted to repro-
gramme the epigenome more extensively and, if also
incorporated into the germ cells, their effects may
become transgenerational1. By contrast, events that
occur outside a sensitive period of brain development
(that is, as an adult) may be more limited in their effect
and duration, as they may produce specific epigenetic
changes that do not manifest in epigenomic or germ cell
reprogramming.
According to the Barker hypothesis6, neurodevelop-
mental reprogramming induces adaptations to the early
environment in anticipation of a similar postnatal envi
ronment. When there is a mismatch between these two
environments, such reprogramming may increase disease
risk. For example, if a mother is exposed to increased
stress during pregnancy but the postpartum environ-
ment is not stressful, the offspring’s brain may continue to
respond inappropriately to stressors that are experienced
throughout life6.
Recent reviews in this area have focused on the spe-
cific developmental periods (including prenatal, postnatal
and pubertal periods) during which such reprogramming
occurs, the types of stimuli that are involved (including
specific diets and exposure to infection or to stress) and
specific epigenetic outcomes2–5,7–9. The goal of this Review
is to integrate these points and highlight potential shared
mechanisms that are important in reprogramming the
developing brain. I examine results from human and
animal studies that shed light on the factors that direct
and shape brain development, and I discuss how differing
parental contributions may affect offspring disease risk
and resilience. In particular, I examine transgenerational
epigenetic effects on neurodevelopment and discuss the
interactions between specific perturbations, develop-
mental windows of susceptibility and the sex-specificity
of these effects.

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 REVIEWS

Environmental Maternal milieu Fetus The importance of the prenatal period. The best evidence
exposures • Cytokines Epigenetic programming of that the timing of insults is a factor in disease risk comes
• Maternal diet • Lipids germ cells and somatic
• Maternal stress • Stress hormones tissue: methyl donors (folate from birth cohort studies examining the effects of maternal
• Maternal infection • Metabolic indices and choline) broadly affect (prepartum) exposure to specific stress or immune chal-
(glucose, insulin the methylome or specific lenges. These studies compile birth registries that enable
and FFAs) imprinted genes
prospective studies to be carried out and thus may have
access to more accurate records of the exposure under
investigation than do retrospective studies. They also
allow the collection of plasma and tissue samples, which
enables potential biomarkers and genetic and/or epigenetic
factors associated with susceptibility to be assessed. For
example, birth cohort studies have revealed strong asso-
ciations between in utero exposure to infection, hypoxia
or starvation and an increased risk of schizophrenia22,23,
as well as an association with impairments in executive
function in patients with schizophrenia15, which indicates
that prenatal brain development, not surprisingly, is a
Placenta uniquely susceptible period. Furthermore, recent stud-
• Cytokines ies showed marked changes in cortical layer patterning,
• Nutrient transporters and receptors which is a prenatal process, in children with ASD, suggest-
(folate and choline)
• Lipid metabolites ing that a disturbance occurred before birth24. However,
• Growth factors (IGF and IGFBP) in‑depth neuro­developmental assessments that compare
• Oxygen perfusion similar perturbations across these prenatal and postnatal
• Glycogen
periods will be necessary to identify a specific period of
development as a crucial factor in disease risk.
Nature Reviews | Neuroscience
Figure 1 | Complex interactions between the maternal milieu, placenta and fetal There is growing evidence that there are specific devel-
compartments during gestation. Fetal antecedents such as maternal dietary opmental windows of increased vulnerability throughout
deficiencies, chronic stress or infection can promote endocrine disruptions in the pregnancy itself 10,25–27. Indeed, a recent epidemiological
maternal milieu, including increases in pro-inflammatory cytokines and stress hormones, study reported an association between maternal stress
and shifts in metabolic indices. In addition, these environmental exposures can indirectly that is experienced specifically during the first trimes-
alter placental development and function by changing local energy metabolism and lipid
ter of pregnancy and an increased risk of schizophrenia
storage and metabolism. This can alter the transmission of key nutrients that are
important for fetal growth and brain development, including growth factors and methyl in male offspring 28. Exposure to maternal infection and
donor nutrients such as folate and choline, which can affect fetal somatic and germ cell high levels of pro-inflammatory cytokines during early or
epigenetic programming. FFAs, free fatty acids; IGF, insulin-like growth factor; IGFBP, mid-gestation was also associated with an increased risk
IGF-binding protein. of neurodevelopmental disorders18. For example, children
of women who were exposed to influenza during early or
mid-pregnancy, but not during late gestation, had a three-
Developmental windows of susceptibility fold higher risk of schizophrenia than those of women
Studies evaluating susceptibility factors in neuro­ who had not been exposed13. Similarly, studies using the
developmental disorders have indicated that certain Danish medical birth register reported that maternal viral
outcomes are commonly linked to exposure to differ- infection resulting in hospitalization in the first trimester,
ent types of gestational and early-life environments. For but not later in pregnancy, was associated with a three-
Birth cohort studies example, exposure to stress or infection during either of fold higher risk of ASD in the child29,30. Furthermore, the
Longitudinal prospective these developmental periods is linked to an increased Early Markers for Autism study found increased levels of
studies in which pregnancies
risk of depression, schizophrenia and autism spectrum 17 cytokines in early and mid-gestation in the mothers
are followed through birth and
into childhood to correlate the disorders (ASDs)1,10–15. This suggests that these insults of children that were later diagnosed with ASD31. These
risk for given outcomes with may promote common mechanisms irrespective of the studies broadly suggest that early pregnancy is a particu-
specified experiences or time at which they occurred in development. However, larly sensitive period in development, during which time
exposures during gestation. epidemiological evidence also supports the idea that insults such as stress and inflammation can promote pro-
These cohorts of individuals
differ from other
the time of exposure during development may be gramming changes that will increase the risk of disease.
epidemiological studies in important for some types of environmental challenge, This increased vulnerability may lie in the sensitivity of
which the outcome measures as similar perturbations occurring at earlier or later the brain, the reactivity of the mother in early gestation
are mainly retrospective. stages have been associated with different long-term and/or the ability of the fetus to recover from such insults.
outcomes1,16–18. Prepubertal and pubertal brain matu- In addition to stress or infection during pregnancy,
Biomarkers
Specific biochemical, ration has been less well examined; however, these reduced maternal nutrition can also have a negative
molecular, anatomical or periods also seem to be susceptible to environmental influence on fetal growth and can induce epigenetic
physiological characteristics disturbances. This is especially true for the prefrontal changes, thereby influencing fetal brain development.
that are used to predict, cortex and its crucial connections within the limbic Long-term epidemiological studies investigating fam-
measure or indicate the
presence or progress of
system, in which disruptions have been linked to an ine exposures during specified periods of pregnancy or
disease or the effects of increased risk of attention deficit hyperactivity disorder postnatal life are beginning to discern associated risks for
treatment. (ADHD) and schizophrenia19–21. neuro­psychiatric disease in first- and second-generation

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REVIEWS
Box 1 | Defining epigenetics and epigenomics
The definition of the term ‘epigenetic’ has been subject to continuous evolution.
Waddington172 coined the term to describe the premise that genes can respond to a
dynamic environment and thereby shape phenotypes such that they are acutely
advantageous to the individual (as opposed to advantageous on a much longer
evolutionary scale). A more recent operational definition narrowed the usage to
include the necessity of heritability, either across generations or to daughter cells,
which involves the erasure and subsequent replacement of epigenetic marks173.
However, over the past decade, developmental and behavioural neuroscientists have
used the term ‘epigenetic modifications’ to refer to functionally relevant chromatin
modifications that alter gene expression but that do not involve a change in DNA
sequence. Recent studies have produced mechanistic insight both at the level of
epigenetics, which examines changes in the expression of specific genes or loci that
result from the addition of epigenetic marks, such as DNA methylation, or from
changes in the levels of a given microRNA, and at the level of epigenomics, which
examines genome-wide changes in gene expression that can be attributed to broad
chromatin modifications. Such outcomes, which might be directed by a given histone
modification (such as histone H3 lysine 4 (H3K4) acetylation) or by changes in the
methylome, can influence large sets of genes that are involved in specific functions
and that generate changes in developmental programming in response to an insult or
a perturbation9.
offspring. Therefore, these studies have the potential to
uncover transgenerational epigenetic mechanisms as well
as direct effects on neurodevelopment. Initial studies have
reported an increased risk of neurodevelopmental disor-
ders in children of women who were exposed to severe
Executive function caloric restriction or famine during pregnancy compared
A set of cognitive abilities that with other periods of postnatal development 32,33. For
include inhibition (resisting instance, children of women who were pregnant during
habits, temptations or the Dutch ‘hunger winter’ of 1944–1945 were more likely
distractions), switching
(adjusting to change) and
to have altered stress responsiveness and schizophrenia
working memory (mentally as adults32,34. These outcomes were also associated with
holding and using information). changes in circulating growth factors and altered DNA
methylation of several genes, including the maternally
Stress responsiveness
imprinted gene insulin-like growth factor 2 (IGF2), which
Behavioural or physiological
measures in response to a were measured 60 years after the famine exposure35. In
stress provocation. In all addition, their second-generation offspring had increased
mammals, the physiological adiposity and overall worse health as adults32. These
hypothalamic–pituitary– transgenerational outcomes suggest that the initial mater-
adrenal stress axis is the
standard measure of stress
nal dietary restriction induced epigenetic marks in fetal
experience. Behavioural germ cells and thus the first-generation offspring were
changes in response to stress able to transmit a phenotype without re‑exposure to the
or threat can also be examined insult. Of course, epidemiological studies are limited by
as an indication of stress state.
an inability to control for the effects of additional mater-
Imprinted gene nal or paternal lifetime experiences and/or interactions
A gene expressed from only with the maternal milieu in subsequent generations. In
one allele in a manner that addition, it is important to note that famine experience
depends on the parent of is also an enormous physiological and psychological
origin and that is regulated by
DNA methylation.
stressor, and stress insults are also metabolic challenges.
Therefore, it is not surprising that both maternal stress
Hypothalamic–pituitary– and maternal famine early in pregnancy produce similar
adrenal stress axis neurodevelopmental disorder risk13–15,32.
(HPA stress axis). The
The current obesity epidemic in Western society has
neuroendocrine core of the
stress system. Its activation shifted the focus from understanding the effect of under-
results in the release of cortico- nutrition to examining the effects of maternal overnutri-
tropin-releasing factor from the tion on neurodevelopment. Indeed, it has been proposed
hypothalamus, adrenocortico- that transgenerational epigenetic mechanisms may con-
tropic hormone from the
pituitary and cortisol
tribute to the rapid rise in obesity rates1. Maternal diet
(corticosterone in rats and and body composition are less dynamic factors than stress
mice) from the adrenal glands. or infection and are therefore more difficult to designate
334 | JUNE 2015 | VOLUME 16
© 2015 Macmillan Publishers Limited. All rights reserved
to a specific period of gestation or even to be limited to
the prenatal environment. However, maternal body mass
and obesity preceding and during pregnancy have been
linked to severity in childhood ADHD symptoms and
other executive function deficits36,37. Exposure to mater-
nal obesity during gestation has also been associated with
a twofold–threefold increased risk of schizophrenia in
offspring in two separate birth cohorts32,33.
As is clear from the findings described above, gesta-
tion is one of the most studied developmental windows
of vulnerability 38,39 and is a critical time in neuro­
developmental programming 13–15,24,32. In this context, it
is important to consider that early pregnancy involves
processes such as embryo implantation and placental dif-
ferentiation, whereas mid- and late gestation are charac-
terized by somatic growth, including considerable brain
development 40. Therefore, exposure to environmental
challenges will influence different developmental pro-
cesses depending on when in pregnancy the exposure
occurred. In addition, the primordial germ cells, which
will give rise to the next generation, undergo remethyla-
tion of imprinted genes before birth in males, whereas
this occurs postnatally in females40. Therefore, if an insult
occurs during a period in which the germ cells are under-
going this normal epigenetic process, changes could be
incorporated that would give rise to a phenotype in future
generations.
Mechanisms of reprogramming
Animal models have been used in an attempt to mimic
the epidemiological findings described above by exam-
ining offspring outcomes that result from specific types
of insults during specific periods of development. These
studies have provided an insight into the mechanisms that
are involved in reprogramming the brain. Rodent brain
development begins during gestation but mostly contin-
ues postnatally. Therefore, using these animal models,
researchers have been able to examine offspring pheno-
types that are relevant to neurodevelopmental disorders
by studying maternal exposure to stress, dietary chal-
lenges and infection both during pregnancy as well as in
the early postnatal period.
Prenatal maternal stress. Studies in which the pregnant
dam is exposed to perturbations that provoke activation
of her neuroendocrine stress system have shown that the
effect of maternal stress during pregnancy on offspring
outcomes, including on stress sensitivity, reward and
cognition, depends on the gestational timing of the expo-
sure (as well as on the sex of the fetus)41–47. In many spe-
cies, including mice, rats, guinea pigs and non-human
primates, maternal stress increases offspring stress
sensitivity such that the overall production of gluco­
corticoids following hypothalamic–pituitary–adrenal stress
axis (HPA stress axis) activation is increased, behav-
ioural stress responses are heightened and cognitive
deficits are present, all of which are endophenotypes
that are associated with neuropsychiatric disease42,46,48–56.
To compare the effects of maternal stress experi-
ence across early, mid- or late pregnancy on the pro-
gramming of offspring stress outcomes, studies in
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Endophenotypes
Quantifiable phenotypes with
an assumed intermediate role
in the pathway from genes to
complex phenotypes. It is
thought that the action of an
endophenotype is easier to
understand biologically and
genetically than the action of
the complex phenotype of
primary interest. They enable
the examination of specific
aspects of complex human
diseases in animal models.
Barnes maze
A spatial learning and memory
task in which the animal is
placed on a large, open table
that has holes around the
circumference. Only one of the
holes has an escape tunnel,
and the animal must learn to
use distal cues to identify this
hole and escape in the shortest
possible time. Learning in this
task involves the hippocampus.
NATURE REVIEWS | NEUROSCIENCE
mice and guinea pigs have used a chronic stress para-
digm42,46,49,50. In mice, exposure to stress very early in
pregnancy (embryonic days 1–7) resulted in male, but
not female, offspring that showed increased immo-
bility in both tail suspension and forced swim tests
as well as cognitive deficits in spatial learning and
memory tasks as adults42,50. Early stress exposure also
affected the programming of stress neurocircuitry:
male offspring showed increased corticosterone pro-
duction in response to acute restraint stress, and this
was associated with increased levels of corticotropin-
releasing factor (CRF) in the amygdala and reduced
hippocampal gluco­c orticoid receptor expression 42.
These changes were associated with corresponding
alterations in promoter methylation, which suggests
that the prenatal stress experience resulted in long-
term epigenetic reprogramming of genes involved in
the development of stress neurocircuitry. The timing
specificity of the effects of prenatal stress was simi-
larly explored using short periods of daily flashing
light as a maternal stressor in guinea pigs at mid- or
late gestation49. Adult offspring that were exposed to
stress in mid-gestation showed increased anxiety-like
behaviours, increased basal corticosterone levels and
lower hippocampal gluco­corticoid receptor expres-
sion; however, late-gestation stress did not produce
these outcomes. This evidence for specific effects of
maternal stress during mid-gestation complements
the evidence arising from the epidemiological studies
described above28,47,49.
Interestingly, in many studies, male offspring are
the most severely affected sex. They often present with
a ‘dysmasculinized’ phenotype; that is, the prenatally
stressed males seem to be more similar to control
females than to control males in terms of their behav-
iours, physiology and gene expression patterns. For
instance, male mice exposed to maternal stress very
early in gestation showed significant changes in spatial
learning and memory performance in a Barnes maze and
used ‘female-like’ strategies to solve the task50. In addi-
tion, these mice showed dysmasculinized patterns of
stress physiology, behaviour and cognitive performance,
which indicates a potential disruption in normal perina-
tal brain masculinization44,52. Similar dysmasculinizing
results have been reported in prenatally stressed rats and
guinea pigs43,46,47,49,57,58. Interestingly, several recent stud-
ies have reported an association between steroidogenic
activity and testosterone levels in fetuses and autism or
autistic-like traits in young children59, and low serum
testosterone levels have been associated with certain
schizophrenia symptoms such as reduced verbal and
working memory in males60. Fetal testosterone levels
also predict the area of the sexually dimorphic grey mat-
ter in the human brain61, and male patients with schizo-
phrenia have orbitofrontal cortex-to-amygdala volume
ratios that are more similar to those of control women
than to those of control men62. These findings support
the idea that brain masculinization is an important
part of normal brain development in males and sug-
gest that this process may be a point of vulnerability to
perturbations in the environment.
© 2015 Macmillan Publishers Limited. All rights reserved
REVIEWS
Postnatal maternal stress. Complex interactions between
the neonate and its mother occur in the postnatal period,
and maternal behaviours such as nursing and grooming
can strongly influence neurodevelopment 63–77. Rodent
studies have shown that both enhanced and fragmented
maternal care in the early postnatal period can result in
marked differences in the behaviours of the adult off-
spring. It is well established that the quality of maternal
care influences epigenetic processes in the developing
brain8,73,78. This was first shown in a rat model in which
dams showed consistent high or low levels of maternal
care — identified as increased or decreased licking and
grooming — and this has since been demonstrated in
many additional species, including mice, hamsters and
non-human primates65–67,70,73,76,79–82. These studies have
shown that the mother’s interactions with her offspring
predict their behavioural and hormonal responses to
stress in adulthood. Specifically, in rats, high rates of
licking and grooming by the mother were associated
with more rapid negative feedback to the HPA stress axis
as a result of enhanced glucocorticoid receptor sensitiv-
ity as well as reduced levels of anxiety-like behaviours72.
At an epigenetic level, these outcomes were correlated
with changes in DNA methylation of the glucocorti-
coid receptor promoter and in glucocorticoid receptor
expression in the hippocampus74. Although other genes
are probably also affected, the glucocorticoid receptor
gene, Nr3c1, seems to have been the most consistently
examined across models and studies. NR3C1 expression
and promoter methylation has also been examined in
humans following early-life trauma, showing a poten-
tially translational outcome for an epigenetic finding
from animal models83,84. However, to adequately address
the question of how maternal stress during pregnancy
ultimately reprogrammes the developing brain, it is nec-
essary to consider the epigenome and the broader epige-
netic mechanisms that would be capable of establishing
changes that are marked enough to alter organismal
behaviour decades later.
Chronic stress during the postnatal period — affecting
the dam and/or the pups — can also affect the cognitive
performance of adult offspring through functional and
structural changes in specific brain regions, including
the hippocampus and the hypothalamus. For example,
in a rat model of altered early postpartum maternal care
resulting from the stress of repeated maternal separation,
the offspring showed lifelong epigenetic changes in the
hippocampus68. Specifically, the ability of a transcrip-
tional repressor to bind to Crf was decreased because of
increased DNA methylation, and this was associated with
a reduced ability of the offspring to appropriately respond
to and to cope during stressful experiences as adults. This
shows an interesting epigenetic reprogramming effect in
response to postnatal stress that opposes changes that
have been observed following prenatal stress: that is, the
Crf promoter was hypomethylated following postna-
tal stress42. This again highlights the importance of the
timing of environmental exposures in producing their
long-term consequences. Interestingly, similar long-term
changes in the expression of CRF in the hypothalamus
were also found in offspring in a model of fragmented
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Microglia
Glia of mesodermal origin and
the resident macrophages of
the CNS.
336 | JUNE 2015 | VOLUME 16
maternal care, in which the rat dam is given limited and
insufficient material with which to build her nest, sug-
gesting that alterations in maternal behaviour during
early postnatal life can drive similar epigenetic events in
her offspring 65,70.
Maternal nutrition and obesity. Rodent studies have
shown a link between prenatal availability of nutrients
and offspring brain development 85. In mice, females
receiving a protein-deficient diet throughout preg-
nancy and lactation produced offspring with intrauter-
ine growth restriction and marked changes in dopamine
circuitry and reward-related behaviours as adults86.
Interestingly, the dopamine reward neurocircuitry was
also affected when pregnant dams were fed a high-fat
diet throughout pregnancy and lactation, which suggests
that the developing reward system is particularly sensi-
tive to gestational nutrition (probably stemming from
an evolutionary benefit of linking motivational behav-
iours with caloric need)87–92. Reduced reward sensitivity
(anhedonia) is a symptom of depression in humans; thus,
there may be a connection between prenatal dietary per-
turbations and an increased risk of affective disorders or
addictions. As described above, maternal undernutrition
also increases stress responsivity in pregnant females and
thus contributes to offspring phenotypes that are simi-
lar to those produced by maternal stress93. For example,
rats that were exposed to 50% food restriction during
pregnancy had increased plasma corticosterone levels
and their adult offspring had higher adrenal weights94.
These maternally food-restricted offspring also showed
changes in the expression of key stress genes, includ-
ing reduced levels of hippocampal Nr3c1 expression.
Furthermore, exposure of female guinea pigs to nutri-
ent restriction for 2 days in the last third of pregnancy
increased maternal and fetal cortisol levels and reduced
the expression of glucocorticoid receptor in the offspring
hypothalamus and hippocampus in adulthood95. These
outcomes are similar to those reported in studies inves-
tigating the effects of prenatal stress and maternal dexa-
methasone treatment 1,22,42,55,56,94,96–98, which suggests that
common signals are produced in the maternal milieu
during maternal stress and undernutrition. These com-
mon signals are probably related to the similar catabolic
metabolism that occurs during these experiences93.
Maternal dietary challenges also affect the maternal
and fetal immune systems. For instance, increases in the
number of hypothalamic microglia and pro-inflamma-
tory cytokine levels were found in fetuses of macaque
mothers who received a high-fat diet before and during
pregnancy 99. As juveniles, the female, but not the male,
offspring showed increased anxiety-like behaviours100.
Similarly, increased numbers of activated microglia in
the hippocampus were reported for the adult offspring
of rat dams fed a diet high in saturated fat throughout
pregnancy and lactation and these animals also showed
increased anxiety-like behaviours and deficits in spa-
tial learning 101. Together, these studies indicate that
there are sex-specific interactions between the mater-
nal diet during early life and long-term neuroimmune
programming 100,101.
© 2015 Macmillan Publishers Limited. All rights reserved
The placenta may function as an important interme-
diary between an unhealthy maternal milieu and fetal
markers of neuroinflammation by producing its own
pro-inflammatory cytokines. Placentas from pregnant
macaques receiving a high-fat diet throughout preg-
nancy showed increased levels of the pro-inflammatory
cytokines tumour necrosis factor (TNF) and inter-
leukin‑6 (IL‑6), and of Toll-like receptor 4 (REF. 102).
Surprisingly, in pregnant rats with restricted access
to low-protein food, levels of TNF and IL‑6 were also
increased in placental tissue103. Timing also seems to be
important for the interaction between the immune sys-
tem and the placenta, as maternal immune activation
during early gestation in mice can result in implanta-
tion failure104,105, and mid-gestation inflammation in
the placenta has been linked to schizophrenia-like and
ASD-like phenotypes106.
Maternal infection and immune activation. Animal
studies examining the effects of maternal infection
during pregnancy have reported many behavioural
outcomes in the adult offspring that are similar to
those detected in offspring exposed to prenatal stress,
including increased stress reactivity and impaired cog-
nitive performance11,12,18,107. This suggests that the two
manipulations may affect the fetus and the developing
brain through common mediators, including glucocor-
ticoids and/or pro-inflammatory cytokines96,108–112. In
fact, inhibition of inflammatory processes in a mouse
model of maternal chronic stress ameliorated the effects
of stress on the offspring 108. The pro-inflammatory
cytokines produced by the mother and/or in the placenta
during maternal infection or chronic stress can elicit
their long-term effects on fetal somatic cells through
interactions with epigenetic machinery. For example,
IL‑6 increases the expression and the activity of DNA
(cytosine 5)-methyltransferase 1 by regulating its gene
promoter 113. In another example, in a mouse model of
early pregnancy stress, changes in the expression of the
chromatin-remodelling enzyme O‑GlcNAc transferase
(OGT) were detected in male but not in female placental
tissue and were found to regulate a broad pattern of gene
expression that is associated with the active histone mark
H3K4me3 (histone H3 lysine 4 trimethylation)38,114.
Transgenerational programming
Although some environmental factors only ‘programme’
the individual that is directly exposed to them, others
are transmitted across one or more generations115. The
transmission of epigenetic marks to future generations
through germ cell reprogramming adds complexity to
our attempts to establish causal links between environ-
mental risk factors and disease. True transgenerational
reprogramming requires an epigenetic mark to be erased
and then re‑established in the germ cell in the absence
of re‑exposure or continuation of the exposure115 (FIG. 2).
Tight control of epigenetic regulation is crucial during
the early phase of gestation, which is characterized by
a wave of genome demethylation followed by de novo
remethylation, establishment of imprints and sex deter-
mination, and is therefore particularly sensitive to
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F1 F2 F3
a Terminal somatic
programming in utero
b Somatic and primordial
germ cell programming
in utero
c Stable germline
inheritance
d Transgenerational
perpetuation of
maternal exposure
e Transgenerational
perpetuation of
paternal exposure
Shows phenotype No phenotype
Nature Reviews | Neuroscience
Figure 2 | Modes of maternal and paternal transgenerational epigenetic
transmission. Several mechanisms for transgenerational epigenetic transmission are
possible, as shown in the table, in which a red mouse indicates that the phenotype is
shown in a particular generation. a | In utero somatic programming may affect tissue and
brain development in the fetus, initiating a developmental trajectory that results in an
adult phenotype in the first (F1) generation. However, this phenotype is not transmitted
to the next generation, as there is no germ cell involvement. b | If in utero somatic
programming is accompanied by changes in primordial germ cells, the phenotype will be
transmitted to a second (F2) generation. In order for this to occur, germ cells must be
exposed to a maternal insult during gestation. c | If the in utero exposure is able to alter
epigenetic marks in the germ line of the F1 offspring and this reprogramming is able to
withstand erasure and re‑establishment during subsequent generations’ germ cell
development, the phenotype will be perpetuated across generations without the need
for re‑exposure to the original insult. This is defined as a truly transgenerational
epigenetic programming. d | Transgenerational phenotypes may also occur through a
repeated maternal effect by which programming during gestation or through exposure
to altered maternal behaviours results in an adult phenotype that is only found in the
maternal lineage. In order for this to occur, the offspring must be re‑exposed to the insult
in each generation to promote the continuation of the phenotype from the dam. This
trait would not be passed through a paternal lineage. e | Transgenerational programming
through a paternal lineage originating from an initial exposure in the male requires
establishment or reprogramming of epigenetic marks in the sperm. Adapted with
permission from REF. 115, Elsevier.
maternal influences that could result in embryonic and
germ cell reprogramming 40. The different ways in which
germ cell epigenetic marks are transmitted and inherited
can also be examined; these include whether the mother
and/or the father pass on the trait and whether male or
female offspring (or both) inherit it. Epidemiological
data from Swedish famine periods suggest that there may
be specific developmental windows during which germ
cells are more vulnerable to this type of environmental
influence116; boys exposed to famine before puberty were
more likely to have children and grandchildren with an
increased risk of cardiovascular disease and diabetes
than males exposed outside this period117. However,
studies have also shown that the type of perturbation or
exposure may be an important determinant in how or if
the germ cells are affected118.
Animal models have been used to examine the
transgenerational outcomes of germ cell reprogram-
ming that are induced by maternal and/or paternal
exposure to an environmental factor. For example, male
mice that are exposed to stress in utero (that is, their
NATURE REVIEWS | NEUROSCIENCE
© 2015 Macmillan Publishers Limited. All rights reserved
REVIEWS
mothers are stressed during pregnancy), which results
in increased stress sensitivity and an increased cortico­
sterone response to acute restraint stress as adults, pass
on this phenotype to their male, but not their female,
offspring 119. Similarly, when mice were exposed to early
postnatal stress (chronic and unpredictable maternal
separation from postnatal days 1 to 14), their stress-
sensitive behavioural phenotype in adulthood contin-
ued through to the third generation120. Changes were
detected in DNA methylation in the germ line of this
third generation, which suggests that germ cell epi­
genetic marks can be maintained across generations
without re‑exposure to the insult 120. These studies sup-
port the idea that the germ line can be manipulated, and
they show the potential ability of external influences to
increase susceptibility or resilience to disease in subse-
quent generations. It is important to note that an envi-
ronmental signal that can influence the germ line does
not necessarily produce a notable phenotype in the first
generation if there is no effect on somatic tissues (includ-
ing the brain). Whether such an effect occurs probably
depends on the timing, the type or the severity of the
exposure121. However, in such a scenario, if the germ cell
were reprogrammed this would give rise to a phenotype
in the second generation; thus, phenotypes can skip a
generation, making it more difficult to form clear risk
analyses for pregnant women or for other at‑risk groups
(FIG. 3). As this premise has not yet been mechanistically
examined, the specific signals or insults remain to be
determined.
Maternal transmission. Studies in which female mice
were exposed to nutritional challenges have provided
some of the clearest examples of how phenotypes can be
lost or retained across generations121–126. These studies
have monitored body length, adiposity, glucose regu-
lation, metabolic rate and behaviour through at least
three generations and have reported epigenetic changes,
including effects on imprinted genes. Maternal obe-
sity or exposure to a high-fat diet during pregnancy is
associated with reduced insulin and glucose sensitiv-
ity and increased body size and adiposity in the first-
generation offspring of humans, non-human primates
and rodents122,127–131. However, not all of these traits will
arise in the second generation, which supports the idea
that many phenotypes are the result of direct somatic cell
programming and not of altered epigenetic marks in the
germ cell. For example, one study in mice found that an
increased offspring body size was the only phenotype of
many from the first generation that was retained into the
second and the third generations following the initial
exposure to a maternal high-fat diet during pregnancy 123.
A closer examination of the transmission pattern of this
phenotype revealed that only second-generation fathers
could pass on the trait and only to their third-generation
daughters, indicating an epigenetic mechanism (as male
mice were not permitted any interaction with the litter,
any passage of traits to their offspring must have been
programmed in their germ cells) involving a pater-
nally expressed gene (as only the second-generation
fathers were able to pass on the trait) that may be on the
VOLUME 16 | JUNE 2015 | 337
REVIEWS
Pregnant female (F0)
Somatic tissue
(including brain)
Placental
contribution
Maternal Fetus (F1)
stress
and diet
Primordial
germ cell (F2)
Figure 3 | Programming of phenotypes and disease risk can skip generations.
Maternal stress or dietary insults can generate signals, in the form of changes in the
hormonal milieu, that are transmitted from the maternalNature Reviews
to the fetal | Neuroscience
compartment via the
placenta. The resulting maternal programming of fetal somatic tissues can lead to
changes in long-term health outcomes in the first generation (F1). Furthermore, in utero
exposure can also occur for the primordial germ cells that will give rise to the second
generation (F2), as these germ cells are also present and undergo reprogramming during
embryonic development. If the exposure does not produce a detectable effect on
somatic tissues but is still able to reprogramme epigenetic marks in the germ cell, a
phenotype could effectively arise in the F2 generation that was not present in the F1
generation, thus having the appearance of ‘skipping a generation’. Adapted with
permission from REF. 115, Elsevier.
X chromosome (as only their daughters inherited the
trait)123. Indeed, the expression pattern of imprinted genes
from these females supported this hypothesis: paternally
expressed genes showed increased overall expression in
the livers of third-generation female offspring born to the
high-fat diet lineage second-generation males but not in
those born to the females.
Paternal transmission. Although transgenerational
effects of various changes in the maternal environment
are well characterized, fewer studies have examined pos-
sible paternal modes of transmission. In such studies,
male mice or rats exposed to social defeat, chronic stress,
cocaine, a high-fat diet, endocrine disrupters or olfactory
Social defeat cues before breeding have been examined132–139. These
A behavioural model in which models provide unique opportunities to examine mech-
rodents (predominantly males) anisms of transgenerational transmission. In a research
are repeatedly exposed to a
laboratory environment, male rodents do not normally
more aggressive conspecific.
The outcomes of these participate in the rearing of their offspring and thus have
confrontations are repeated no postpartum behavioural influence on their develop-
losses for the subject. This ment. Therefore, the ability of the sire to transmit a trait
exposure is then followed by a to his offspring is limited to the genetic and the epige-
period of housing the defeated
animal in close proximity to the
netic material in his sperm, which is an accessible and
aggressor to establish the plentiful tissue source for epigenetic analyses. Therefore,
conditioning. paternal models provide a more transparent method by
338 | JUNE 2015 | VOLUME 16
© 2015 Macmillan Publishers Limited. All rights reserved
which to study mechanisms of epigenetic transmis-
sion than models of maternal transmission, which are
complicated by the potentially confounding effects
of maternal behaviours, parturition and lactation. As
stated above, the term ‘transgenerational’ requires that
an epigenetic mark giving rise to a phenotype is found
in the germ cell and is passed on to the next generation
in the absence of re‑exposure. Therefore, to be consid-
ered a transgenerational epigenetic mechanism in a male
model of transmission, a phenotype need be detected
only into the second generation. Perturbations that
occur during pregnancy, such as maternal stress or infec-
tion, must affect the third generation for their influence
to be truly transgenerational, because the gestating first-
generation offspring and the germ cells that will give rise
to the second-generation offspring are simultaneously
exposed to the initial perturbation115 (FIG. 2).
Epigenetic marks normally found in sperm include
changes in the expression of non-coding RNAs (micro-
RNAs (mi­RNAs) and PIWI-interacting RNAs (piRNAs;
which are important in silencing retrotransposons)),
histone modifications (for the few retained histones in
sperm) and DNA methylation140. Perturbations such
as stress can alter these marks, resulting in offspring
with detectable phenotypes. For instance, when male
mice were exposed to chronic social defeat or chronic
variable stress before breeding, their offspring showed
behavioural and physiological phenotypes of stress
dysregulation, including altered HPA axis responses
to stress132,134. Furthermore, one study showed that,
in sperm from fathers who were chronically stressed
either during puberty or as adults, the levels of a sub-
set of specific mi­RNAs were increased134. Interestingly,
rewarding or reinforcing environmental factors, such as
drugs of abuse, pleasant odours or high-fat diets, can
also produce germline epigenetic marks that are trans-
mitted to the offspring. For example, male rats exposed
to a specific odour paired with a shock passed this
conditioned experience to their offspring through an
epigenetic germ cell mechanism133. Similarly, male off-
spring of male rats that were exposed to chronic cocaine
self-administration showed reward neurocircuitry
hypofunction136. This phenotype was associated with a
change in histone H3 acetylation at the brain-derived
neurotrophic factor (Bdnf) promoter in the sperm of the
sires that were initially exposed, and the same change
was also detected in the brains of their male offspring.
In another study, female but not male offspring of male
rats that were exposed to a high-fat diet for 10 weeks
had reduced glucose sensitivity and β-cell dysfunction135.
Such sex-specific outcomes suggest that the epigenetic
mark or machinery involved is found on the X or the
Y chromosome and/or that the offspring phenotype is
dependent on programming or activational effects of
oestradiol or testosterone. The mechanism by which
paternal life experiences such as stress, diets or drugs can
reprogramme epigenetic marks in the germ cell are not
currently understood; however, normal processes and
development across spermatogenesis and the stages at
which epigenetic or transcriptional machinery is present
in the cell must be considered (FIG. 4).
www.nature.com/reviews/neuro
 REVIEWS

Epididymis

Window of vulnerability for programming and reprogramming epigenetic marks during spermatogenesis

Active transcription Meiotic Active transcription Chromatin

divisions compaction

RNA storage, RNA-binding proteins and translational repression

Chromatid body

Spermatogonium Early Late 1st meiotic 2nd Round Step 8 round Elongating Final sperm
spermatocyte spermatocyte division spermatocyte spermatid spermatid spermatid maturation

Figure 4 | Windows of vulnerability to environmental reprogramming in spermatogenesis. Nature The figure| Neuroscience
Reviews shows the
stages of spermatogenesis and sperm maturation, highlighting the periods in which environmental exposures can
reprogramme epigenetic marks. Active transcription and storage of RNA is ongoing through the spermatogonium,
spermatocyte and spermatid stages. However, once compaction of paternal DNA has occurred, which involves replacing
the majority of histones with protamines, there is not a clear mechanism by which epigenetic marks can be further altered
in the mature sperm. This suggests that an environmental exposure must occur within a sensitive time period of
spermatogenesis in order to pass on a new or reprogrammed epigenetic mark to future offspring. In most mammals,
complete spermatogenesis occurs in 6–8 weeks, by which point sperm are present in the epididymis for final maturation
processes. The time between chromatin compaction and the transit of mature sperm to the epididymis is at least 10 days,
during which time epigenetic and transcriptional machineries are no longer active. However, the secretion of exosomes
and other factors that interact with the maturing sperm in the epididymis may impart lasting epigenetic marks, such as
microRNAs. Figure is reproduced from Bale T. Lifetime stress experience: transgenerational epigenetics and germ cell
programming. Dialogues Clin. Neurosci. 2014;16:297–305 (REF. 179), with the permission of © AICH-Servier Research
Group, Suresnes, France, and adapted from REF. 180, Nature Publishing Group.

Pre-pulse inhibition
A reduction in the magnitude
of the startle reflex that occurs
when an organism is presented
with a non-startling stimulus (a
pre-pulse) before being
presented with the startling
stimulus. Deficits in pre-pulse
inhibition have been observed
in patients with schizophrenia.
Sex specificity in epigenetic mechanisms
As described above, epigenetic responses to environmen-
tal changes can be sex specific at several levels. The per-
ception or the response to the exposure itself can differ
between sexes, so that one sex produces a higher or a lower
change in hormone levels or greater neural activation,
leading to a cascade of differing outcomes. The placenta
is an example of a tissue that shows sex-specific effects;
for example, maternal stress or dietary challenge during
pregnancy produces lasting gene expression changes in
the placentas of male fetuses but not in the placentas of
female fetuses38,39,42. This provides a mechanism by which
the effects of stress can be transmitted in a sex-specific
manner to the developing brain. Epidemiological stud-
ies have shown that gender is an important determinant
in the severity and the onset of diseases that are associ-
ated with certain fetal antecedents. For example, male,
but not female, children of women who were in their
second trimester of pregnancy during the 1940 invasion
of the Netherlands (and therefore who presumably suf-
fered high stress levels at that time) had an increased risk
of schizophrenia141. A recent report found that maternal
depression during pregnancy correlated with increased
postnatal anxiety development in children and that males
were significantly more likely to be affected142. Many neu-
rodevelopmental disorders, such as ASD, have a strong
sex bias143. Exposure to maternal stress before week 32 of
gestation has been suggested to be a contributing factor
to ASD144. Therefore, studies examining the role of fetal
sex in determining the specific outcomes of maternal
stress during pregnancy may provide a unique insight
into neurodevelopmental disease aetiologies145.
There is also evidence that the effects of activation
of the maternal immune system on the offspring are
sex dependent. For example, male offspring of preg-
nant rats exposed to lipopolysaccharides late in gesta-
tion showed deficits in pre-pulse inhibition in adulthood,
whereas female offspring were unaffected146. Similarly,
male, but not female, mice exposed during late gestation
to maternal injection of the viral mimic polyinosinic–
polycytidylic acid (poly(I:C)) showed behavioural and
cognitive inflexibility in adulthood, resembling some of
the negative symptoms of schizophrenia18. These effects
were also associated with reduced levels of glutamate in
the prefrontal cortex in males. Such studies support the
widely held view that males are more vulnerable to pre-
natal insults than females, who are more susceptible to
insults that occur in the postnatal environment 147.
Animal studies have also begun to examine the
developmental timing of such sex-specific outcomes
more closely1. For example, as described above, gesta-
tion has been established as a period in which important
masculinization of the male brain can be disrupted by
maternal stress22,42,50,57. Disruptions in the organizational
programming of the sexually dimorphic brain may also
have a role in the aetiology of neurodevelopmental dis-
orders. For example, structural brain volume analyses
showed a reduction of the normal sexual dimorphism of
the brain in male patients with schizophrenia62,148. These
studies are exciting because they suggest a specific time
window in development (probably prenatal) in which
an exposure or insult disrupted this normal masculin-
izing process. As the female brain is not exposed to
gonadal hormones during this early developmental

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REVIEWS

Box 2 | Common cellular mechanisms?

In thinking about the mechanisms by which prenatal perturbations promote transgenerational outcomes, it is interesting
to note that several seemingly different insults, such as maternal stress, diet and infection, can produce similar
neurodevelopmental and phenotypic outcomes. Mechanistically, this suggests that there are a limited number of
physiological signals from the maternal milieu that are responsible for transmitting information from the in utero
environment to the developing fetus and that physiological disturbances produced by maternal stress, infection, famine
or obesity share some common end points.
The most evolutionarily important signals are likely to be related to energy availability, which is important in
determining future fitness, and are therefore likely to include metabolic signals such as insulin, glucose, free fatty acids
and glucocorticoids, as well as inflammatory cytokines (FIG. 1). Glucocorticoids are certainly an important downstream
indicator of stress, and some aspects of prenatal stress models can be recapitulated by simply administering high levels of
glucocorticoids to the pregnant animal55,56,96,174,175. However, the stress response encompasses other stress hormones,
such as catecholamines, as well as numerous downstream metabolic factors. The involvement of these additional
metabolic processes in developmental programming at the level of the placenta and the fetus is probable and has been
shown to be important in rodent models of prenatal stress38,93,97,114,176.
There can only be a limited number of biochemical inputs that a given cell can to respond to and, as such, it is likely that
different environmental challenges impinge on the same signalling cascades and downstream effectors to promote
changes in existing epigenetic marks or to apply new ones — effectively reprogramming similar offspring phenotypes
from different starting points. Therefore, there must be a cellular barometer by which the quantitative and the qualitative
changes in the hormonal milieu are surveyed. For instance, during maternal or paternal stress exposure, for a signal to be
gauged as important enough to warrant activation of the epigenetic machinery to facilitate the creation of new or
changed epigenetic marks, does a stressor need to be of a certain duration, to reach a certain level (perceived as mild
versus severe or chronic) or to induce a certain level of stress hormones? How does a germ cell know whether a stressor
warrants passing on the information to future generations? Coincident detection of key signals occurring over precise
periods is probably required.

period, these findings indicate a sex-specific process
that is a point of potential disruption in males. This
may ultimately lead to a mismatch for these males later
in life, as they would be genetically XY but would not
have a fully masculinized brain.
Gonadal hormones and their receptors also interact
with epigenetic mechanisms to promote normal brain
maturation and development149. For instance, oestradiol
can affect epigenetic machinery, including DNA methyl­
transferases and histone deacetylases (HDACs), in the
neonate as a part of sexually dimorphic brain develop-
ment, and can alter DNA methylation patterns3,150–152.
Therefore, alterations in the levels of oestrogen, or its
precursor testosterone, that result from perturbations
experienced during early life can manifest as long-term
reprogramming of brain function149. Oestradiol can also
alter histone modification through actions on HDACs149.
A single postnatal administration of an HDAC inhibitor
in male mice was shown to disrupt the sexual dimorphic
development of the bed nucleus of the stria terminalis
(BNST), which is a crucial part of the neurocircuitry
involved in regulating stress responses153. Small non-
coding mi­RNAs also seem to be responsive to oestradiol
levels in the early postnatal mouse brain; more than 250
of the most abundantly expressed mi­RNAs in the brain
significantly differ in their expression between males
and females119. These sex differences are dependent on
the conversion of gonadal testosterone to oestradiol in
males154, as a single administration of an aromatase inhibi-
tor at birth was able to completely shift the entire male
miRNA expression pattern to that of a female119.
These studies show that gonadal hormones can,
during important periods, promote epigenomic effects
and thereby broadly influence neurodevelopment,
effectively producing the sexually dimorphic brain.
Importantly, these regulatory mechanisms can be sensi-
tive to perturbations in the environment. As described
above, maternal stress and infection are both associated
with dysmasculinized phenotypes, which suggests that
they cause a disruption in male sexual differentiation
or gonad development that in turn alters testosterone
production or its actions in the brain during this critical
neonatal organizational period1,155,156. Finally, it is also
interesting to note that although the Y chromosome con-
tains no miRNA genes, the mammalian X chromosome
is highly enriched for them and their density is approxi-
mately twofold higher than it is on autosomes in mice
and humans147. In circumstances in which these genes
escape X inactivation, females would have twice the
expression level of males, thus producing a sex-specific
post-transcriptional mode of regulation that provides
females with the select ability to buffer or to respond
differently to insults.
Conclusions and future directions
It is clear that the outcomes and the long-term sustain-
ability of dynamic epigenetic marks are dependent on
factors such as the timing and the type of exposure,
the sex of the offspring and, in the case of transgenera-
tional effects, the sex of the parent and the offspring.
According to the Barker hypothesis, whether such
programming ultimately results in disease risk or resil-
ience mainly depends on the concordance between the
expected and actual environments. As discussed above,
there are several common signalling mechanisms
through which perturbations during early develop-
ment are likely to function, including inflammation and
energy metabolism in the maternal milieu and in the
placenta (BOX 2). Studies examining the early postnatal
environment have also identified common epigenetic

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© 2015 Macmillan Publishers Limited. All rights reserved

 REVIEWS

Box 3 | Caution in translation of epigenetic outcomes

and ultimate brain function. Studies in mice focus-
ing on brain region-specific epigenetic processes and
We currently lack the ability to accurately predict who will present with a important behaviours related to cognitive performance,
neuropsychiatric disease. However, the collection of biological samples as part of learning and memory, and appetitive behaviours have
prospective studies may enable us to identify predictive biomarkers, which may include repeatedly shown how dynamic the epigenome contin-
epigenetic marks, and to distinguish between disease cause and effect if changes in a
ues to be into adulthood1,9,69,160–170. This reminds us that
given outcome precede disease onset rather than being a response to it. Of course, this
raises the question of whether epigenetic and epigenomic changes in peripheral cells
insults that occur early in life may be just a ‘first hit,’
and tissues are informative about what goes on in the brain, as most epigenetic changes which primes the brain to be more or less responsive to
are cell and tissue specific177. A great deal more evidence is still needed before we can future insults, with disease presentation occurring fol-
draw firm conclusions as to how accurate the epigenome will be in relaying the story of lowing multiple insults in life. Support for this hypoth-
an individual’s life exposures and experiences and our ability to translate that into esis can be seen in neuropsychiatric diseases in which
predictions for disease risk and resilience. adult stress-induced onset occurs, such as depression
Using established epidemiological and clinical measures to design better animal or post-traumatic stress disorder. Perturbations experi-
models that reproduce specific endophenotypes rather than attempting to model enced during neurodevelopment that promote dysregu-
mental health disorders in totality is an important goal that may facilitate study of the lation of stress pathways would certainly be considered
translational potential of the epigenome. Such models may enable us, for example, to
an important first hit 12,22,171.
better understand how an environmental factor may programme susceptibility or
resilience to disease in humans. However, if the field of transgenerational epigenetics
The field of transgenerational epigenetics has been
has taught us anything, it is that experience is everything. Results from studies showing on a fast-paced trajectory over the past decade, with
transgenerational effects of stress, odorant exposure, diet and infection have opened the development of animal models that reliably and
our eyes to the dynamic (that is, programmable and reprogrammable) epigenome, predictively mimic epidemiological studies in order to
including — and importantly — the germline epigenome5,133–135,178. The seemingly determine the mechanisms by which perturbations in
standard experiences that our experimental animals encounter, including shipping, the environment are associated with risk of neurode-
dietary changes, parasitic infections and treatments, construction noise and different velopmental disorders (BOX 3). Future studies will need
environments (such as germ-free versus conventional facilities), can produce to use ‘omic’ technologies to better understand the
substantial epigenetic differences within otherwise genetically identical inbred strains. broader reprogramming abilities of the environment
If present in the germ cells, these marks can last for generations, promoting phenotypic
on the epigenome. For instance, metabolomic analyses
variability within and between research laboratories. Controlling for such variables is
difficult but important.
of the maternal and the fetal milieu would be useful in
models of maternal stress or infection to determine the
metabolic signals that are involved in neurodevelop-
gene targets in the brain. In addition, recent studies have ment. There remain several considerable gaps in our
provided insight into the novel contributions of the gut knowledge: what are the physiological and the cellular
microbiome to early brain development; thus, metabolic signals by which experience is able to reprogramme
and immune processes may interact to ultimately affect an epigenetic mark (BOX 1), and when and how are the
neurodevelopment 157–159. Taken together with other germ cells involved? Identifying these mechanisms in a
important gene targets, these findings will provide the stepwise manner will facilitate the discovery of better
clues necessary to discover the upstream transcriptional disease predictive biomarkers and will perhaps lead to
regulatory mechanisms, which will increase our under- effective interventions for disease prevention and earlier
standing of the links between developmental experience treatments (BOX 3).

1. Bale, T. L. et al. Early life programming and
neurodevelopmental disorders. Biol. Psychiatry 68,
314–319 (2010).
2. Curley, J. P., Jensen, C. L., Mashoodh, R. &
Champagne, F. A. Social influences on neurobiology
and behavior: epigenetic effects during development.
Psychoneuroendocrinology 36, 352–371 (2011).
3. McCarthy, M. M. & Nugent, B. M. Epigenetic
contributions to hormonally mediated sexual
differentiation of the brain. J. Neuroendocrinol.
25,1133–1140 (2013).
4. Hodes, G. E. Sex, stress, and epigenetics: regulation of
behavior in animal models of mood disorders. Biol.
Sex Differ. 4, 1 (2013).
5. Bohacek, J., Gapp, K., Saab, B. J. & Mansuy, I. M.
Transgenerational epigenetic effects on brain
functions. Biol. Psychiatry 73, 313–320 (2013).
6. Hales, C. N. & Barker, D. J. The thrifty phenotype
hypothesis. Br. Med. Bull. 60, 5–20 (2001).
In this paper, the thrifty phenotype hypothesis
describes how a mismatch between fetal
programming and the postnatal environment can
result in disease risk.
7. Binder, E. B. & Nemeroff, C. B. The CRF system, stress,
depression and anxiety-insights from human genetic
studies. Mol. Psychiatry 15, 574–588 (2010).
8. Hackman, D. A., Farah, M. J. & Meaney, M. J.
Socioeconomic status and the brain: mechanistic
insights from human and animal research. Nature Rev.
Neurosci. 11, 651–659 (2010).
9. Sweatt, J. D. The emerging field of neuroepigenetics.
Neuron 80, 624–632 (2013).
10. Glynn, L. M., Wadhwa, P. D., Dunkel-Schetter, C.,
Chicz-Demet, A. & Sandman, C. A. When stress
happens matters: effects of earthquake timing on
stress responsivity in pregnancy. Am. J. Obstet.
Gynecol. 184, 637–642 (2001).
11. Deverman, B. E. & Patterson, P. H. Cytokines and CNS
development. Neuron 64, 61–78 (2009).
12. Howerton, C. L. & Bale, T. L. Prenatal programing: at
the intersection of maternal stress and immune
activation. Horm. Behav. 62, 237–242 (2012).
13. Brown, A. S. Epidemiologic studies of exposure to
prenatal infection and risk of schizophrenia and
autism. Dev. Neurobiol. 72, 1272–1276 (2012).
14. Brown, A. S. et al. Prenatal infection and cavum
septum pellucidum in adult schizophrenia. Schizophr.
Res. 108, 285–287 (2009).
15. Brown, A. S. et al. Prenatal exposure to maternal
infection and executive dysfunction in adult
schizophrenia. Am. J. Psychiatry 166, 683–690
(2009).
16. Selevan, S. G., Kimmel, C. A. & Mendola, P. Identifying
critical windows of exposure for children’s health.
Environ. Health Perspect. 108 (Suppl. 3), 451–455
(2000).
17. Rapoport, J. L., Addington, A. M., Frangou, S. &
Psych, M. R. The neurodevelopmental model of
schizophrenia: update 2005. Mol. Psychiatry 10,
434–449 (2005).
18. Meyer, U., Feldon, J. & Dammann, O. Schizophrenia
and autism: both shared and disorder-specific
pathogenesis via perinatal inflammation? Pediatr. Res.
69, 26R–33R (2011).
This study examines the shared mechanistic link
between neurodevelopmental disorders that are
related to prenatal inflammation.
19. Morrison, K. E., Rodgers, A. B., Morgan, C. P. &
Bale, T. L. Epigenetic mechanisms in pubertal brain
maturation. Neuroscience 264, 7–24 (2013).
20. Davis, E. P., Sandman, C. A., Buss, C., Wing, D. A. &
Head, K. Fetal glucocorticoid exposure is associated
with preadolescent brain development. Biol.
Psychiatry 74, 647–655 (2013).
21. Malter Cohen, M. et al. Early-life stress has persistent
effects on amygdala function and development in mice
and humans. Proc. Natl Acad. Sci. USA 110,
18274–18278 (2013).
22. Bale, T. L. Sex differences in prenatal epigenetic
programming of stress pathways. Stress 14, 348–356
(2011).
23. Susser, E., St Clair, D. & He, L. Latent effects of prenatal
malnutrition on adult health: the example of schizophrenia.
Ann. NY Acad. Sci. 1136, 185–192 (2008).
24. Stoner, R. et al. Patches of disorganization in the
neocortex of children with autism. N. Engl. J. Med.
370, 1209–1219 (2014).
25. Avishai-Eliner, S., Brunson, K. L., Sandman, C. A. &
Baram, T. Z. Stressed-out, or in (utero)? Trends
Neurosci. 25, 518–524 (2002).

NATURE REVIEWS | NEUROSCIENCE VOLUME 16 | JUNE 2015 | 341

© 2015 Macmillan Publishers Limited. All rights reserved

REVIEWS
26. Wadhwa, P. D., Sandman, C. A. & Garite, T. J. The
neurobiology of stress in human pregnancy:
implications for prematurity and development of the
fetal central nervous system. Prog. Brain Res. 133,
131–142 (2001).
27. Wadhwa, P. D., Sandman, C. A., Porto, M.,
Dunkel-Schetter, C. & Garite, T. J. The association
between prenatal stress and infant birth weight and
gestational age at birth: a prospective investigation.
Am. J. Obstet. Gynecol. 169, 858–865 (1993).
28. Khashan, A. S. et al. Higher risk of offspring
schizophrenia following antenatal maternal exposure
to severe adverse life events. Arch. Gen. Psychiatry
65, 146–152 (2008).
29. Atladottir, H. O. et al. Association of hospitalization for
infection in childhood with diagnosis of autism
spectrum disorders: a Danish cohort study. Arch.
Pediatr. Adolesc. Med. 164, 470–477 (2010).
30. Atladottir, H. O. et al. Maternal infection requiring
hospitalization during pregnancy and autism spectrum
disorders. J. Autism Dev. Disord. 40, 1423–1430
(2010).
31. Goines, P. E. et al. Increased midgestational IFNγ, IL‑4
and IL‑5 in women bearing a child with autism: a case-
control study. Mol. Autism 2, 13 (2011).
32. Brown, A. S. & Susser, E. S. Prenatal nutritional
deficiency and risk of adult schizophrenia. Schizophr.
Bull. 34, 1054–1063 (2008).
33. Xu, M. Q. et al. Prenatal malnutrition and adult
schizophrenia: further evidence from the 1959–1961
Chinese famine. Schizophr. Bull. 35, 568–576
(2009).
34. Ravelli, G. P., Stein, Z. A. & Susser, M. W. Obesity in
young men after famine exposure in utero and early
infancy. N. Engl. J. Med. 295, 349–353 (1976).
35. Heijmans, B. T. et al. Persistent epigenetic differences
associated with prenatal exposure to famine in
humans. Proc. Natl Acad. Sci. USA 105,
17046–17049 (2008).
36. Van Lieshout, R. J., Taylor, V. H. & Boyle, M. H.
Pre-pregnancy and pregnancy obesity and
neurodevelopmental outcomes in offspring:
a systematic review. Obes. Rev. 12, e548–e559
(2011).
37. Buss, C. et al. Impaired executive function mediates
the association between maternal pre-pregnancy body
mass index and child ADHD symptoms. PLoS ONE 7,
e37758 (2012).
38. Howerton, C. L., Morgan, C. P., Fischer, D. B. &
Bale, T. L. O‑GlcNAc transferase (OGT) as a placental
biomarker of maternal stress and reprogramming of
CNS gene transcription in development. Proc. Natl
Acad. Sci. USA 110, 5169–5174 (2013).
39. Mao, J. et al. Contrasting effects of different maternal
diets on sexually dimorphic gene expression in the
murine placenta. Proc. Natl Acad. Sci. USA 107,
5557–5562 (2010).
40. Weaver, J. R., Susiarjo, M. & Bartolomei, M. S.
Imprinting and epigenetic changes in the early
embryo. Mamm. Genome 20, 532–543 (2009).
41. Mueller, B. R. & Bale, T. L. Impact of prenatal stress on
long term body weight is dependent on timing and
maternal sensitivity. Physiol. Behav. 88, 605–614
(2006).
42. Mueller, B. R. & Bale, T. L. Sex-specific programming
of offspring emotionality after stress early in
pregnancy. J. Neurosci. 28, 9055–9065 (2008).
This study identifies sex-specific changes in the
placenta in response to early gestational stress
that are associated with changes in offspring
neurodevelopment.
43. Brunton, P. J. & Russell, J. A. Prenatal social stress in
the rat programmes neuroendocrine and behavioural
responses to stress in the adult offspring: sex-specific
effects. J. Neuroendocrinol. 22, 258–271 (2010).
44. Brunton, P. J. & Russell, J. A. Allopregnanolone and
suppressed hypothalamo–pituitary–adrenal axis
stress responses in late pregnancy in the rat. Stress
14, 6–12 (2011).
45. Weinstock, M. Gender differences in the effects of
prenatal stress on brain development and behaviour.
Neurochem. Res. 32, 1730–1740 (2007).
46. Kapoor, A., Kostaki, A., Janus, C. & Matthews, S. G.
The effects of prenatal stress on learning in adult
offspring is dependent on the timing of the stressor.
Behav. Brain Res. 197, 144–149 (2009).
47. Kapoor, A., Leen, J. & Matthews, S. G. Molecular
regulation of the hypothalamic–pituitary–adrenal axis
in adult male guinea pigs after prenatal stress at
different stages of gestation. J. Physiol. 586,
4317–4326 (2008).
342 | JUNE 2015 | VOLUME 16
48. Schneider, M. L., Moore, C. F., Kraemer, G. W.,
Roberts, A. D. & DeJesus, O. T. The impact of prenatal
stress, fetal alcohol exposure, or both on
development: perspectives from a primate model.
Psychoneuroendocrinology 27, 285–298 (2002).
49. Kapoor, A. & Matthews, S. G. Short periods of
prenatal stress affect growth, behaviour and
hypothalamo–pituitary–adrenal axis activity in male
guinea pig offspring. J. Physiol. 566, 967–977
(2005).
50. Mueller, B. R. & Bale, T. L. Early prenatal stress impact
on coping strategies and learning performance is sex
dependent. Physiol. Behav. 91, 55–65 (2007).
51. Lemaire, V., Koehl, M., Le Moal, M. & Abrous, D. N.
Prenatal stress produces learning deficits associated
with an inhibition of neurogenesis in the hippocampus.
Proc. Natl Acad. Sci. USA 97, 11032–11037 (2000).
52. Darnaudery, M. & Maccari, S. Epigenetic
programming of the stress response in male and
female rats by prenatal restraint stress. Brain Res.
Rev. 57, 571–585 (2008).
53. Weinstock, M. Alterations induced by gestational
stress in brain morphology and behaviour of the
offspring. Prog. Neurobiol. 65, 427–451 (2001).
54. Coe, C. L. et al. Prenatal stress diminishes
neurogenesis in the dentate gyrus of juvenile rhesus
monkeys. Biol. Psychiatry 54, 1025–1034 (2003).
55. Welberg, L. A., Seckl, J. R. & Holmes, M. C. Prenatal
glucocorticoid programming of brain corticosteroid
receptors and corticotrophin-releasing hormone:
possible implications for behaviour. Neuroscience
104, 71–79 (2001).
56. Welberg, L. A. & Seckl, J. R. Prenatal stress,
glucocorticoids and the programming of the brain.
J. Neuroendocrinol. 13, 113–128 (2001).
57. Ward, I. L. Prenatal stress feminizes and
demasculinizes the behavior of males. Science 175,
82–84 (1972).
This is one of the first studies to determine the
importance of sex-specific neurodevelopmental
programming that can be disrupted by maternal
stress.
58. Ward, I. L. & Stehm, K. E. Prenatal stress feminizes
juvenile play patterns in male rats. Physiol. Behav. 50,
601–605 (1991).
59. Baron-Cohen, S. et al. Elevated fetal steroidogenic
activity in autism. Mol. Psychiatry 20, 369–376
(2015).
60. Moore, L. et al. Serum testosterone levels are related
to cognitive function in men with schizophrenia.
Psychoneuroendocrinology 38, 1717–1728 (2013).
61. Lombardo, M. V. et al. Fetal testosterone influences
sexually dimorphic gray matter in the human brain.
J. Neurosci. 32, 674–680 (2012).
62. Gur, R. E. et al. A sexually dimorphic ratio of
orbitofrontal to amygdala volume is altered in
schizophrenia. Biol. Psychiatry 55, 512–517 (2004).
This paper reports the dysmasculinization of limbic
brain regions in men with schizophrenia.
63. Fenoglio, K. A. et al. Enduring, handling-evoked
enhancement of hippocampal memory function and
glucocorticoid receptor expression involves activation
of the corticotropin-releasing factor type 1 receptor.
Endocrinology 146, 4090–4096 (2005).
64. Fenoglio, K. A., Brunson, K. L. & Baram, T. Z.
Hippocampal neuroplasticity induced by early-life
stress: functional and molecular aspects. Front.
Neuroendocrinol. 27, 180–192 (2006).
65. Ivy, A. S., Brunson, K. L., Sandman, C. & Baram, T. Z.
Dysfunctional nurturing behavior in rat dams with
limited access to nesting material: a clinically relevant
model for early-life stress. Neuroscience 154,
1132–1142 (2008).
66. Ivy, A. S. et al. Hippocampal dysfunction and cognitive
impairments provoked by chronic early-life stress
involve excessive activation of CRH receptors.
J. Neurosci. 30, 13005–13015 (2010).
67. Korosi, A. & Baram, T. Z. Plasticity of the stress
response early in life: mechanisms and significance.
Dev. Psychobiol. 52, 661–670 (2010).
68. Korosi, A. et al. Early-life experience reduces
excitation to stress-responsive hypothalamic neurons
and reprograms the expression of corticotropin-
releasing hormone. J. Neurosci. 30, 703–713 (2010).
69. McClelland, S., Korosi, A., Cope, J., Ivy, A. &
Baram, T. Z. Emerging roles of epigenetic mechanisms
in the enduring effects of early-life stress and
experience on learning and memory. Neurobiol. Learn.
Mem. 96, 79–88 (2011).
70. Rice, C. J., Sandman, C. A., Lenjavi, M. R. &
Baram, T. Z. A novel mouse model for acute and long-
© 2015 Macmillan Publishers Limited. All rights reserved
lasting consequences of early life stress. Endocrinology
149, 4892–4900 (2008).
This novel and reproducible model shows how
disruptions in maternal care can manifest in
reprogramming of offspring stress pathways in the
brain.
71. Ladd, C. O. et al. Long-term behavioral and
neuroendocrine adaptations to adverse early
experience. Prog. Brain Res. 122, 81–103 (2000).
72. Liu, D. et al. Maternal care, hippocampal
glucocorticoid receptors, and hypothalamic–pituitary–
adrenal responses to stress. Science 277,
1659–1662 (1997).
73. Meaney, M. J. Maternal care, gene expression, and
the transmission of individual differences in stress
reactivity across generations. Annu. Rev. Neurosci. 24,
1161–1192 (2001).
74. Weaver, I. C. et al. Epigenetic programming by
maternal behavior. Nature Neurosci. 7, 847–854
(2004).
This important paper was the first to establish
epigenetic mechanisms in the developing brain that
were altered by the quality of maternal care.
75. Barha, C. K., Pawluski, J. L. & Galea, L. A. Maternal
care affects male and female offspring working
memory and stress reactivity. Physiol. Behav. 92,
939–950 (2007).
76. Sanchez, M. M., Ladd, C. O. & Plotsky, P. M. Early
adverse experience as a developmental risk factor for
later psychopathology: evidence from rodent and
primate models. Dev. Psychopathol. 13, 419–449
(2001).
77. Caldji, C. et al. Maternal care during infancy regulates
the development of neural systems mediating the
expression of fearfulness in the rat. Proc. Natl Acad.
Sci. USA 95, 5335–5340 (1998).
78. Weaver, I. C., Diorio, J., Seckl, J. R., Szyf, M. &
Meaney, M. J. Early environmental regulation of
hippocampal glucocorticoid receptor gene expression:
characterization of intracellular mediators and
potential genomic target sites. Ann. NY Acad. Sci.
1024, 182–212 (2004).
79. Korosi, A. & Baram, T. Z. The pathways from mother’s
love to baby’s future. Front. Behav. Neurosci. 3, 27
(2009).
80. Graham, Y. P., Heim, C., Goodman, S. H., Miller, A. H.
& Nemeroff, C. B. The effects of neonatal stress on
brain development: implications for psychopathology.
Dev. Psychopathol. 11, 545–565 (1999).
81. Lyons, D. M., Martel, F. L., Levine, S., Risch, N. J. &
Schatzberg, A. F. Postnatal experiences and genetic
effects on squirrel monkey social affinities and
emotional distress. Horm. Behav. 36, 266–275
(1999).
82. Barr, C. S. et al. Sexual dichotomy of an interaction
between early adversity and the serotonin transporter
gene promoter variant in rhesus macaques. Proc. Natl
Acad. Sci. USA 101, 12358–12363 (2004).
83. McGowan, P. O. et al. Epigenetic regulation of the
glucocorticoid receptor in human brain associates with
childhood abuse. Nature Neurosci. 12, 342–348
(2009).
84. Vukojevic, V. et al. Epigenetic modification of the
glucocorticoid receptor gene is linked to traumatic
memory and post-traumatic stress disorder risk in
genocide survivors. J. Neurosci. 34, 10274–10284
(2014).
85. Koshy, T. S., Sara, V. R., King, T. L. & Lazarus, L. The
influence of protein restriction imposed at various
stages of pregnancy on fetal and placental
development. Growth 39, 497–506 (1975).
86. Vucetic, Z. et al. Early life protein restriction alters
dopamine circuitry. Neuroscience 168, 359–370
(2010).
87. Pankevich, D. E., Teegarden, S. L., Hedin, A. D.,
Jensen, C. L. & Bale, T. L. Caloric restriction
experience reprograms stress and orexigenic
pathways and promotes binge eating. J. Neurosci. 30,
16399–16407 (2010).
88. Teegarden, S. L. & Bale, T. L. Decreases in dietary
preference produce increased emotionality and risk for
dietary relapse. Biol. Psychiatry 61, 1021–1029
(2007).
89. Teegarden, S. L. & Bale, T. L. Effects of stress on
dietary preference and intake are dependent on access
and stress sensitivity. Physiol. Behav. 93, 713–723
(2008).
90. Teegarden, S. L., Scott, A. N. & Bale, T. L. Early life
exposure to a high fat diet promotes long-term
changes in dietary preferences and central reward
signaling. Neuroscience 162, 924–932 (2009).
www.nature.com/reviews/neuro

91. Will, M. J., Franzblau, E. B. & Kelley, A. E. Nucleus
accumbens μ‑opioids regulate intake of a high-fat diet
via activation of a distributed brain network.
J. Neurosci. 23, 2882–2888 (2003).
92. Pecina, S., Cagniard, B., Berridge, K. C., Aldridge, J. W.
& Zhuang, X. Hyperdopaminergic mutant mice have
higher “wanting” but not “liking” for sweet rewards.
J. Neurosci. 23, 9395–9402 (2003).
93. Cottrell, E. C., Holmes, M. C., Livingstone, D. E.,
Kenyon, C. J. & Seckl, J. R. Reconciling the nutritional
and glucocorticoid hypotheses of fetal programming.
FASEB J. 26, 1866–1874 (2012).
94. Lesage, J., Blondeau, B., Grino, M., Breant, B. &
Dupouy, J. P. Maternal undernutrition during late
gestation induces fetal overexposure to glucocorticoids
and intrauterine growth retardation, and disturbs the
hypothalamo–pituitary–adrenal axis in the newborn
rat. Endocrinology 142, 1692–1702 (2001).
95. Lingas, R., Dean, F. & Matthews, S. G. Maternal
nutrient restriction (48 h) modifies brain corticosteroid
receptor expression and endocrine function in the fetal
guinea pig. Brain Res. 846, 236–242 (1999).
96. Cottrell, E. C. & Seckl, J. R. Prenatal stress,
glucocorticoids and the programming of adult disease.
Front. Behav. Neurosci. 3, 19 (2009).
97. Seckl, J. R. & Holmes, M. C. Mechanisms of disease:
glucocorticoids, their placental metabolism and fetal
‘programming’ of adult pathophysiology. Nature Clin.
Pract. Endocrinol. Metab. 3, 479–488 (2007).
98. Welberg, L. A., Thrivikraman, K. V. & Plotsky, P. M.
Chronic maternal stress inhibits the capacity to
up‑regulate placental 11β‑hydroxysteroid
dehydrogenase type 2 activity. J. Endocrinol. 186,
R7–R12 (2005).
99. Grayson, B. E. et al. Changes in melanocortin
expression and inflammatory pathways in fetal
offspring of nonhuman primates fed a high-fat diet.
Endocrinology 151, 1622–1632 (2010).
100. Sullivan, E. L. et al. Chronic consumption of a high-fat
diet during pregnancy causes perturbations in the
serotonergic system and increased anxiety-like
behavior in nonhuman primate offspring. J. Neurosci.
30, 3826–3830 (2010).
101. Bilbo, S. D. & Tsang, V. Enduring consequences of
maternal obesity for brain inflammation and behavior
of offspring. FASEB J. 24, 2104–2115 (2010).
102. Frias, A. E. et al. Maternal high-fat diet disturbs
uteroplacental hemodynamics and increases the
frequency of stillbirth in a nonhuman primate model of
excess nutrition. Endocrinology 152, 2456–2464
(2011).
103. Shen, Q. et al. The role of pro-inflammatory factors in
mediating the effects on the fetus of prenatal
undernutrition: implications for schizophrenia.
Schizophr. Res. 99, 48–55 (2008).
104. Ilievski, V., Lu, S. J. & Hirsch, E. Activation of toll-like
receptors 2 or 3 and preterm delivery in the mouse.
Reprod. Sci. 14, 315–320 (2007).
105. Hsiao, E. Y. & Patterson, P. H. Placental regulation of
maternal–fetal interactions and brain development.
Dev. Neurobiol. 72,1317–1326 (2012).
106. Malkova, N. V., Yu, C. Z., Hsiao, E. Y., Moore, M. J. &
Patterson, P. H. Maternal immune activation yields
offspring displaying mouse versions of the three core
symptoms of autism. Brain Behav. Immun. 26,
607–616 (2012).
107. Bilbo, S. D. & Schwarz, J. M. Early-life programming of
later-life brain and behavior: a critical role for the
immune system. Front. Behav. Neurosci. 3, 14 (2009).
108. Bronson, S. L. & Bale, T. L. Prenatal stress-induced
increases in placental inflammation and offspring
hyperactivity are male-specific and ameliorated by
maternal antiinflammatory treatment. Endocrinology
155, 2635–2646 (2014).
109. Hsiao, E. Y. & Patterson, P. H. Activation of the
maternal immune system induces endocrine changes
in the placenta via IL‑6. Brain Behav. Immun. 25,
604–615 (2011).
110. Lucassen, P. J. et al. Prenatal stress reduces postnatal
neurogenesis in rats selectively bred for high, but not
low, anxiety: possible key role of placental
11β‑hydroxysteroid dehydrogenase type 2. Eur.
J. Neurosci. 29, 97–103 (2009).
111. Mairesse, J. et al. Maternal stress alters endocrine
function of the feto-placental unit in rats. Am.
J. Physiol. Endocrinol. Metab. 292, E1526–E1533
(2007).
112. Garbett, K. A., Hsiao, E. Y., Kalman, S.,
Patterson, P. H. & Mirnics, K. Effects of maternal
immune activation on gene expression patterns in the
fetal brain. Transl. Psychiatry 2, e98 (2012).
NATURE REVIEWS | NEUROSCIENCE
113. Hodge, D. R. et al. Interleukin‑6 regulation of the
human DNA methyltransferase (HDNMT) gene in
human erythroleukemia cells. J. Biol. Chem. 276,
39508–39511 (2001).
114. Howerton, C. L. & Bale, T. L. Targeted placental
deletion of OGT recapitulates the prenatal stress
phenotype including hypothalamic mitochondrial
dysfunction. Proc. Natl Acad. Sci. USA 111,
9639–9644 (2014).
115. Dunn, G. A., Morgan, C. P. & Bale, T. L. Sex-specificity
in transgenerational epigenetic programming. Horm.
Behav. 59, 290–295 (2011).
116. Pembrey, M. E. et al. Sex-specific, male-line
transgenerational responses in humans. Eur. J. Hum.
Genet. 14, 159–166 (2006).
117. Kaati, G., Bygren, L. O. & Edvinsson, S. Cardiovascular
and diabetes mortality determined by nutrition during
parents’ and grandparents’ slow growth period. Eur.
J. Hum. Genet. 10, 682–688 (2002).
118. Pembrey, M., Saffery, R. & Bygren, L. O. Human
transgenerational responses to early-life experience:
potential impact on development, health and
biomedical research. J. Med. Genet. 51, 563–572
(2014).
This is a comprehensive review that covers the field
of transgenerational epigenetics in relation to
human health and disease.
119. Morgan, C. P. & Bale, T. L. Early prenatal stress
epigenetically programs dysmasculinization in second-
generation offspring via the paternal lineage.
J. Neurosci. 31, 11748–11755 (2011).
120. Franklin, T. B. et al. Epigenetic transmission of the
impact of early stress across generations. Biol.
Psychiatry 68, 408–415 (2010).
121. King, V. et al. Maternal obesity has little effect on
the immediate offspring but impacts on the next
generation. Endocrinology 154, 2514–2524
(2013).
122. Dunn, G. A. & Bale, T. L. Maternal high-fat diet
promotes body length increases and insulin
insensitivity in second-generation mice. Endocrinology
150, 4999–5009 (2009).
123. Dunn, G. A. & Bale, T. L. Maternal high-fat diet effects
on third-generation female body size via the paternal
lineage. Endocrinology 152, 2228–2236 (2011).
124. Jimenez-Chillaron, J. C. et al. Intergenerational
transmission of glucose intolerance and obesity by
in utero undernutrition in mice. Diabetes 58,
460–468 (2009).
125. Thamotharan, M. et al. Transgenerational inheritance
of the insulin-resistant phenotype in embryo-
transferred intrauterine growth-restricted adult female
rat offspring. Am. J. Physiol. Endocrinol. Metab. 292,
E1270–E1279 (2007).
126. Burdge, G. C. et al. Dietary protein restriction of
pregnant rats in the F0 generation induces altered
methylation of hepatic gene promoters in the adult
male offspring in the F1 and F2 generations. Br. J. Nutr.
97, 435–439 (2007).
127. Boney, C. M., Verma, A., Tucker, R. & Vohr, B. R.
Metabolic syndrome in childhood: association with
birth weight, maternal obesity, and gestational
diabetes mellitus. Pediatrics 115, e290–e296
(2005).
128. McCurdy, C. E. et al. Maternal high-fat diet triggers
lipotoxicity in the fetal livers of nonhuman primates.
J. Clin. Invest. 119, 323–335 (2009).
129. Sullivan, E. L., Smith, M. S. & Grove, K. L. Perinatal
exposure to high-fat diet programs energy balance,
metabolism and behavior in adulthood.
Neuroendocrinology 93, 1–8 (2011).
This paper examines the sex-specific
neurodevelopmental programming and resulting
increased anxiety in the non-human primate that
follows exposure to a maternal high-fat diet during
pregnancy.
130. Tamashiro, K. L., Terrillion, C. E., Hyun, J., Koenig, J. I.
& Moran, T. H. Prenatal stress or high-fat diet
increases susceptibility to diet-induced obesity in rat
offspring. Diabetes 58, 1116–1125 (2009).
131. Shiell, A. W., Campbell, D. M., Hall, M. H. &
Barker, D. J. Diet in late pregnancy and glucose-insulin
metabolism of the offspring 40 years later. BJOG 107,
890–895 (2000).
132. Dietz, D. M. & Nestler, E. J. From father to offspring:
paternal transmission of depressive-like behaviors.
Neuropsychopharmacology 37, 311–312 (2012).
133. Dias, B. G. & Ressler, K. J. Parental olfactory
experience influences behavior and neural structure in
subsequent generations. Naure Neurosci. 17, 89–96
(2014).
© 2015 Macmillan Publishers Limited. All rights reserved
REVIEWS
This is a comprehensive examination, in adult male
mice, of epigenetic changes in germ cells that occur
in response to exposure to an odour cue in fear
conditioning that is related to reprogramming of
offspring brain and behaviour.
134. Rodgers, A. B., Morgan, C. P., Bronson, S. L.,
Revello, S. & Bale, T. L. Paternal stress exposure alters
sperm microRNA content and reprograms offspring
HPA stress axis regulation. J. Neurosci. 33,
9003–9012 (2013).
This paper identifies important changes in specific
sperm mi­RNAs produced by chronic stress in adult
mice that correlate with programming of offspring
stress reactivity.
135. Ng, S. F. et al. Chronic high-fat diet in fathers
programs β-cell dysfunction in female rat offspring.
Nature 467, 963–966 (2010).
136. Vassoler, F. M., White, S. L., Schmidt, H. D.,
Sadri-Vakili, G. & Pierce, R. C. Epigenetic inheritance
of a cocaine-resistance phenotype. Nature Neurosci.
16, 42–47 (2013).
137. Carone, B. R. et al. Paternally induced
transgenerational environmental reprogramming of
metabolic gene expression in mammals. Cell 143,
1084–1096 (2010).
138. Anway, M. D., Cupp, A. S., Uzumcu, M. &
Skinner, M. K. Epigenetic transgenerational actions of
endocrine disruptors and male fertility. Science 308,
1466–1469 (2005).
139. Anderson, L. M. et al. Preconceptional fasting of
fathers alters serum glucose in offspring of mice.
Nutrition 22, 327–331 (2006).
140. Jenkins, T. G. & Carrell, D. T. The sperm epigenome
and potential implications for the developing embryo.
Reproduction 143, 727–734 (2012).
141. van Os, J. & Selten, J. P. Prenatal exposure to
maternal stress and subsequent schizophrenia. The
May 1940 invasion of The Netherlands. Br.
J. Psychiatry 172, 324–326 (1998).
142. Gerardin, P. et al. Depression during pregnancy: is the
developmental impact earlier in boys? A prospective
case-control study. J. Clin. Psychiatry 72, 378–387
(2010).
143. Newschaffer, C. J. et al. The epidemiology of autism
spectrum disorders. Annu. Rev. Publ. Health 28,
235–258 (2007).
144. Beversdorf, D. Q. et al. Timing of prenatal stressors
and autism. J. Autism Dev. Disord. 35, 471–478
(2005).
145. Sandman, C. A., Glynn, L. M. & Davis, E. P. Is there a
viability–vulnerability tradeoff? Sex differences in fetal
programming. J. Psychosomat. Res. 75, 327–335
(2013).
146. Smith, S. E., Li, J., Garbett, K., Mirnics, K. &
Patterson, P. H. Maternal immune activation alters
fetal brain development through interleukin‑6.
J. Neurosci. 27, 10695–10702 (2007).
147. Morgan, C. P. & Bale, T. L. Sex differences in
microRNA regulation of gene expression: no smoke,
just miRs. Biol. Sex Differ. 3, 22 (2012).
148. Cowell, P. E., Kostianovsky, D. J., Gur, R. C.,
Turetsky, B. I. & Gur, R. E. Sex differences in
neuroanatomical and clinical correlations in
schizophrenia. Am. J. Psychiatry 153, 799–805
(1996).
149. McCarthy, M. M. et al. The epigenetics of sex
differences in the brain. J. Neurosci. 29,
12815–12823 (2009).
150. Nugent, B. M. & McCarthy, M. M. Epigenetic
underpinnings of developmental sex differences in the
brain. Neuroendocrinology 93, 150–158 (2011).
151. Westberry, J. M., Trout, A. L. & Wilson, M. E.
Epigenetic regulation of estrogen receptor-α gene
expression in the mouse cortex during early postnatal
development. Endocrinology 151, 731–740 (2010).
152. Kurian, J. R., Olesen, K. M. & Auger, A. P. Sex
differences in epigenetic regulation of the estrogen
receptor-α promoter within the developing preoptic
area. Endocrinology 151, 2297–2305 (2010).
153. Murray, E. K., Hien, A., de Vries, G. J. & Forger, N. G.
Epigenetic control of sexual differentiation of the bed
nucleus of the stria terminalis. Endocrinology 150,
4241–4247 (2009).
154. McCarthy, M. M. Molecular aspects of sexual
differentiation of the rodent brain.
Psychoneuroendocrinology 19, 415–427 (1994).
155. Bale, T. L. Stress sensitivity and the development of
affective disorders. Horm. Behav. 50, 529–533 (2006).
156. Bale, T. L. Neuroendocrine and immune influences on
the CNS: it’s a matter of sex. Neuron 64, 13–16
(2009).
VOLUME 16 | JUNE 2015 | 343
REVIEWS
157. Cryan, J. F. & Dinan, T. G. Mind-altering
microorganisms: the impact of the gut microbiota on
brain and behaviour. Nature Rev. Neurosci. 13,
701–712 (2012).
158. Dinan, T. G. & Cryan, J. F. Regulation of the stress
response by the gut microbiota: implications for
psychoneuroendocrinology. Psychoneuroendocrinology
37, 1369–1378 (2012).
159. Clarke, G. et al. The microbiome–gut–brain axis
during early life regulates the hippocampal
serotonergic system in a sex-dependent manner. Mol.
Psychiatry 18, 666–673 (2013).
160. Borrelli, E., Nestler, E. J., Allis, C. D. &
Sassone-Corsi, P. Decoding the epigenetic language
of neuronal plasticity. Neuron 60, 961–974
(2008).
161. Colvis, C. M. et al. Epigenetic mechanisms and gene
networks in the nervous system. J. Neurosci. 25,
10379–10389 (2005).
162. Tsankova, N., Renthal, W., Kumar, A. & Nestler, E. J.
Epigenetic regulation in psychiatric disorders. Nature
Rev. Neurosci. 8, 355–367 (2007).
163. Day, J. J. et al. DNA methylation regulates associative
reward learning. Nature Neurosci. 16, 1445–1452
(2013).
164. Roth, T. L. & Sweatt, J. D. Regulation of chromatin
structure in memory formation. Curr. Opin. Neurobiol.
19, 336–342 (2009).
165. Sweatt, J. D. Experience-dependent epigenetic
modifications in the central nervous system. Biol.
Psychiatry 65, 191–197 (2009).
344 | JUNE 2015 | VOLUME 16
166. Zovkic, I. B., Guzman-Karlsson, M. C. & Sweatt, J. D.
Epigenetic regulation of memory formation and
maintenance. Learn. Mem. 20, 61–74 (2013).
167. Zovkic, I. B. & Sweatt, J. D. Epigenetic mechanisms in
learned fear: implications for PTSD.
Neuropsychopharmacology 38, 77–93 (2013).
168. Mahgoub, M. & Monteggia, L. M. A role for histone
deacetylases in the cellular and behavioral
mechanisms underlying learning and memory. Learn.
Mem. 21, 564–568 (2014).
169. Adachi, M. & Monteggia, L. M. Decoding transcriptional
repressor complexes in the adult central nervous
system. Neuropharmacology 80, 45–52 (2014).
170. Lester, B. M. et al. Behavioral epigenetics. Ann. NY
Acad. Sci. 1226, 14–33 (2011).
171. Morrison, K. E., Rodgers, A. B., Morgan, C. P. &
Bale, T. L. Epigenetic mechanisms in pubertal brain
maturation. Neuroscience 264, 17–24 (2014).
172. Waddington, C. H. Canalization of development and
genetic assimilation of acquired characters. Nature
183, 1654–1655 (1959).
This paper laid the foundation for the field of epigenetic
programming by defining the ‘epigenetic landscape’.
173. Berger, S. L., Kouzarides, T., Shiekhattar, R. &
Shilatifard, A. An operational definition of epigenetics.
Genes Dev. 23, 781–783 (2009).
174. Drake, A. J., Walker, B. R. & Seckl, J. R.
Intergenerational consequences of fetal programming
by in utero exposure to glucocorticoids in rats. Am.
J. Physiol. Regul. Integr. Comp. Physiol. 288,
R34–R38 (2005).
© 2015 Macmillan Publishers Limited. All rights reserved
175. O’Regan, D., Welberg, L. L., Holmes, M. C. &
Seckl, J. R. Glucocorticoid programming of pituitary–
adrenal function: mechanisms and physiological
consequences. Semin. Neonatol. 6, 319–329 (2001).
176. Nyirenda, M. J., Welberg, L. A. & Seckl, J. R.
Programming hyperglycaemia in the rat through prenatal
exposure to glucocorticoids — fetal effect or maternal
influence? J. Endocrinol. 170, 653–660 (2001).
177. Heller, E. A. et al. Locus-specific epigenetic remodeling
controls addiction- and depression-related behaviors.
Nature Neurosci. 17, 1720–1727 (2014).
178. Brunner, A. M., Nanni, P. & Mansuy, I. M. Epigenetic
marking of sperm by post-translational modification of
histones and protamines. Epigenetics Chromatin 7, 2
(2014).
179. Bale, T. L. Lifetime stress experience:
transgenerational epigenetics and germ cell
programming. Dialogues Clin. Neurosci. 16, 297–305
(2014).
180. Kotaja, N. & Sassone-Corsi, P. The chromatid body:
a germ-cell-specific RNA-processing centre. Nature
Rev. Mol. Cell Biol. 8, 85–90 (2007).
Acknowledgements
This work was supported by grants from the US National
Institutes of Health MH099910, MH104184, MH091258,
MH087597 and MH073030.
Competing interests statement
The author declares no competing interests.
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